Grain Jars Stopped Colonizing

[blue.magic]

I have inoculated several 900 ml rye jars.

Unfortunately, the colonization seems to have stopped in all the jars.

The first picture is the jar inoculated from agar slurry on Nov 11. There are patches of uncolonized rye and it looks the same for the last 10 days.

The second picture is the jar inoc. from liquid culture on Nov 25. I have shaken the jar a week ago to make new inoc. sites but there are few colonized patches and nothing changed.

The third jar is inoculated from spore solution on Nov 12, not shaken. Only the top part got colonized and the rye seems completely dry.

Is there anything I can do to salvage these jars? What could have possibly gone wrong? Has it dried out? The incubator is a clear plastic box (closed with several 6 mm holes on top and bottom of front and back walls) with heating pad below keeping the temperature around 26-27 C (79-80 F) all the time.

There are no signs of contamination, just while fluffy mycelium with occassional rhizomorphic growth. I guess if there were contams, it would have been already colonized by molds.

The rye has been soaked with a bit of gypsum for 12 hours (it started germinating very quickly so I avoided longer soaks). Then boiled for 15 minutes, then strained and dried by shaking the collander for approx. 15 minutes, then loaded in jars and sterilized at 15 psi for 90 minutes, cooled overnigh inside PC and inoculated the next day.

I am not sure about the cooking times - some say to cook the rye for 10 minutes, others say 60 minutes. Same with sterilization - some say 20 minutes are OK but others say 90 minutes...

 

[Nereus]

I was just about to write a long methodology about how to troubleshoot stalled jars but then I've read your post again and noticed that you've already tried many known methods, although there is some method that you haven't said anything about or haven't tried it yet. Gas exchange!

In the pics we can't see how you dialed in your setting in regards to GE, that would have helped. Still you can loosen up a bit on your poly filters, make very small holes in the micropore tape with a sterile needle etc.

Also why are you using an incubator? Are the temps in your house so low that one is mandatory? If the temps are good for you and you can mainly hang around in a t shirt I'd say an incu. is not needed.

Another thing I can think about that can help redistribute humidity inside the jars would be shaking. How many times have you shaken them? I know you can do it twice, even up to three times without hurting the myc. Could it be time for a second shake?

If all of the above have been tried with no success it then means that the issue happened some time during the grain prep i.e insufficient hydration of the sub. With rye I'd go for at least a 24hrs soak. Usually I like a 48hrs one. Germination is low so I don't mind a couple sprouting kernels, they won't survive the simmer and the PC process anyway. Then hydrate on a low simmer until minimal amount of kernels start to burst. That's when you know you've achieved a proper water content. Only PC after you've hydrated well. Do it for 90min not less cause it's better to feel safe about them, about this work in general. Ime if you have guidelines like these that you follow aswell, success is granted 100% of the times.

 

[blue.magic]

Here you can see my setup.

Gas exchange is the best bet since I left the box mostly closed (venting it once every 1-2 days) and made the GE holes in the box just a few days ago.

It's winter here and the temperatures are 17-23 C (65-73 F) so heating is necessary as the temps should be above 24 C (76 F) I think.

The incubator is placed in a storage room that is not heated by the central heating.

I have shaken the jars just once in the beginning as I read on forums that most people don't shake them at all or just once. I will try it then.

Thanks about the tip on simmering kernels. I heard in one video that bursting kernels are bad and should be avoided. But not that it's a good indicator of moisture content.

In my case almost ALL the rye kernels sprouted right the next day, not sure why they went alive so quickly...

Thanks for the guidelines. I have watched many videos, entire "Let's Grow Mushrooms" tutorial and read entire "The Psilocybin Mushroom Bible" but they don't troubleshoot all the problems that occur in practice...

 

[Felnik]

Those first pics of your jars look totally green mold contaminated to me unless I'm missing something .

 

[dmusicaltrancistor]

that first jar is almost colonized if you want to salvage it you may be able to go ahead and spawn it to a bulk sub i have done this is the past with great success just make sure your substrate is properly pasteurized and it should still be all right

 

[Nereus]

Those first pics of your jars look totally green mold contaminated to me unless I'm missing something .

It's just white, very fluffy myc. Light and cam. colours might mislead one to believe it is mold, but it is not.

Looking at the first pics from an angle I'd def. say your moisture is on point. There is some condensation on the walls of the jars too.

Yeah, and about GE, you have to get some in if you want to promote healthy and fast growth. It is known for myc to produce CO2 as it develops. It does not produce alot and it is said to be tolerant to it, but it adds up and it does slow things down some. Loosen up the filters a bit, maintaining a sterile procedure obviously, and they'll explode with growth again after 24hrs.

 

[Nereus]

I have shaken the jars just once in the beginning as I read on forums that most people don't shake them at all or just once. I will try it then.[/i]

I'm all for shaking. I tried both shaking and not shaking and overall I can say that shaken myc is better, stronger and colonises fully and integrally faster than unshaken one.

You say that you've shaken the jars once in the beginning. Was it after inoculation? Or was it after the myc reached a certain colonisation %?

From my experience, jars are best left unshaken for a while after inoculation. Then after the myc reaches something like a solid 30% or more, give it the first shake, then once again after reaching 60-70% give it another and it should be enough. Some even shake one last time before spawning to ensure that the myc is healthy and that colonisation is even, but I find that unnecessary.

Also you might want to let your jars consolidate for a while after they have reached 100% point. It's a good and healthy practice, and it encourages the growth of larger fruiting bodies.

 

[Loveall]

I second the shaking notion. It can help a lot.

I've done split experiments from jars in the same batch with shaking and without shaking. It took a lot longer for the unshaken jars to colonize. I've also never noticed negative effects from shaking.

 

[blue.magic]

One problem is that after I drilled the GE holes in the box, the temperature dropped several degrees and now its barely above the level for colonization, most of the box is under 23 C while the minimum colonization temp is 24 C

I cannot "loosen up" the air holes in the jars. It's a micropore tape glued from both sides over 2 mm hole in the jar lid. Maybe the sharp needle would help but this will certainly make it unsterile - I guess the mycelium will withstand that at this point though. I will drill a bigger hole or multiple small holes for the future jars...

you may be able to go ahead and spawn it to a bulk sub i have done this is the past with great success just make sure your substrate is properly pasteurized and it should still be all right

Okay I will spawn the one of the almost colonized jars and see.

You say that you've shaken the jars once in the beginning. Was it after inoculation? Or was it after the myc reached a certain colonisation %?

It was right after inoculation for the agar ones. The ones inoculated from liquid cultures were shaken 10 days after inoc where it was like 10% colonized, then it colonized a bit more and stays like that since.

Those first pics of your jars look totally green mold contaminated to me unless I'm missing something.

There is no green mold. There was yellowish light, the jar sits on washing mashine that is actually white.
I will use fluorescent tubes later and color correct the photos.

 

[kerelsk]

I think getting your incubator down in tempt a little bit is a good idea since the fungus itself will create heat inside the jar as it breaks down the grain. I think your incubator may have been too warm, thereby evaporating the humidity out of the grains too fast as it is.

There has been discussion on the shroomery as to the best temperature to colonize, and many say that the "ideal temp" of 86F is actually a bit warm in most circumstances. Even the tropical cubensis will grow in conditions of 50-60 degrees F, very slowly, but steadily, and the produced mushrooms are meatier.

And I think you're stalled because there's not enough water in the grains to support rapid growth of the fungus. The fungus will hang on and even slowly colonize, but ultimately fail unless there's more water added.

You could try to add some sterilized water with a syringe into the jars and hope the grain/fungus soaks it up well enough, but it's less than ideal.

I would go ahead and make some jars, perhaps see that the grain is a bit more soaked. 12 hour soak + 15 min boil isn't bad but perhaps just a slightly longer boil? I squeeze the kernels and have a certain hydration level I look for.
I would also make sure the jars don't get above 86F/30C, more in the 70 degree range.

Hope you get to the bottom of this, and your fungus loves your conditions the conditions you're giving it

 

[Nereus]

One problem is that after I drilled the GE holes in the box, the temperature dropped several degrees and now its barely above the level for colonization, most of the box is under 23 C while the minimum colonization temp is 24 C

I cannot "loosen up" the air holes in the jars. It's a micropore tape glued from both sides over 2 mm hole in the jar lid. Maybe the sharp needle would help but this will certainly make it unsterile - I guess the mycelium will withstand that at this point though. I will drill a bigger hole or multiple small holes for the future jars...

I encourage you to take a sterile syringe needle and puncture the micropore tape a couple of times. Very small holes will still not let any airbornes or anything pass through, especially if you keep them enclosed in the incubator too. I used to pierce my tape all the time for more GE and I never had contam. problems because of this reason. On the contrary, it always helped to get a faster move on.

On the temp. subject, I would not worry too much as long as it doesn't go under 21 centigrades. After it starts colonising, mycelium produces heat so it keeps itself warm above 18C (i.e when stored at room temps, ofc this does not apply for cold, very cold or freezing temps) so your 23 degrees is on point. At that temp. I wouldn't expect things to slow down. Because of the warmth of the myc I find an incubator is not needed in general, but this certainly depends alot on the other factors aswell.

 

[blue.magic]

On the temp. subject, I would not worry too much as long as it doesn't go under 21 centigrades.

I have read the optimal temp is close to 30 centigrades but since the mycelium produces heat, something like 27 C is optimal. There are so many opinions on the correct temperature going from 19 to 30 C ... it's very hard to find a solid information.

I don't know the actual temperature since it changes rapidly from center to walls of the incubator.

The jars are even warm to touch on one side and very cold on the other, depending which side is closer to the center of the incubator.

I am thinking about gluing a tin foil to the walls of the incubator box so the heat will even out.

 

[downwardsfromzero]

Once upon a time, my jars did very well incubated in an expanded polystyrene box without any heating once the mycelium started to appear. FAE didn't seem to be a problem either. Lids were alu foil and Tyvek held with just the outer ring of the metal disc lids. Incubation temperature was not measured, or at least not recorded. Should have taken more notes, really. Fruited with a TiT at 26-27°C, without casing because it just happened.

Best of luck!

 

[Nereus]

On the temp. subject, I would not worry too much as long as it doesn't go under 21 centigrades.

I have read the optimal temp is close to 30 centigrades but since the mycelium produces heat, something like 27 C is optimal. There are so many opinions on the correct temperature going from 19 to 30 C ... it's very hard to find a solid information.

I don't know the actual temperature since it changes rapidly from center to walls of the incubator.

The jars are even warm to touch on one side and very cold on the other, depending which side is closer to the center of the incubator.

I am thinking about gluing a tin foil to the walls of the incubator box so the heat will even out.

As stated before by kerelsk, I can second that, rapid colonising mycelium due to higher temps and a more tropical type microclimate will exhibit denser but much smaller fruiting bodies, whereas the slow grower that is also kept under medium warm microclimate will have an end result of fairly dense but larger fruits.
Now it certainly depends on what you are looking for, both sides can have advantages as well as the opposite.
One way to even the temperature of the jars would be to constantly rotate them and interchange their place so they all get to be as warm/chill ast the others, and ofc some insulation won't hurt. What room temperature have you got ?

As a conlusion, the only true wisdom when it comes to these things will only come out of more and more subsequent experimentation. Taking notes on progress helps alot for future reference also and, imo, all the advise one can get will never be enough. Because each individual environment is unique with it's own set of variables, only through constant observation and fine tuning you can reach a sort of baseline, from where you can then always begin and know that you can achieve your goals from that point forward.

 

[blue.magic]

On the temp. subject, I would not worry too much as long as it doesn't go under 21 centigrades.

I have read the optimal temp is close to 30 centigrades but since the mycelium produces heat, something like 27 C is optimal. There are so many opinions on the correct temperature going from 19 to 30 C ... it's very hard to find a solid information.

I don't know the actual temperature since it changes rapidly from center to walls of the incubator.

The jars are even warm to touch on one side and very cold on the other, depending which side is closer to the center of the incubator.

I am thinking about gluing a tin foil to the walls of the incubator box so the heat will even out.

As stated before by kerelsk, I can second that, rapid colonising mycelium due to higher temps and a more tropical type microclimate will exhibit denser but much smaller fruiting bodies, whereas the slow grower that is also kept under medium warm microclimate will have an end result of fairly dense but larger fruits.
Now it certainly depends on what you are looking for, both sides can have advantages as well as the opposite.
One way to even the temperature of the jars would be to constantly rotate them and interchange their place so they all get to be as warm/chill ast the others, and ofc some insulation won't hurt. What room temperature have you got ?

As a conlusion, the only true wisdom when it comes to these things will only come out of more and more subsequent experimentation. Taking notes on progress helps alot for future reference also and, imo, all the advise one can get will never be enough. Because each individual environment is unique with it's own set of variables, only through constant observation and fine tuning you can reach a sort of baseline, from where you can then always begin and know that you can achieve your goals from that point forward.

I agree. I have updated both the incubator and FC many times, slowly improving with every batch, not necessarily following book rules.

The ambient room temp is 20.1° C (68.18° F) and it may drop a degree or two as the winter progresses.

I will try to put another (small) heat pad inside the incubator and set the thermostat to lower temp, say 24-25° C (77° F).

I would like larger and stronger fruit bodies for practical reasons as these are easier to pick and it takes less time.

Once upon a time, my jars did very well incubated in an expanded polystyrene box

Great idea! Polystyrene should make even better insulator.

 

[blue.magic]

Okay here is the pic of incubator upgrade.

1. I have glued some alu foil to the bottom and top to reflect heat
2. added smaller heating mat directly inside the box
3. added some plastic foam to the top and bottom as an insulator and to provide offset from heat source
4. placed wire mesh on the foam to provide stable suppor for jars

The thermometer on bottom level now reads 24.1 C (75.38 F) and the top part of one walls is 22.4 C (72.3 F).

I think these temperatures are okay. The ambient humidity seems to be around 50% (+- 3%) year-round. Maybe I will add more alu foil to other walls to improve temp control and to save energy.

I have also added 10 ml of sterile water to the jars via syringe and no shaking as the jars have been alread shaken.

 

[Amphiprion]

I encourage you to take a sterile syringe needle and puncture the micropore tape a couple of times. Very small holes will still not let any airbornes or anything pass through, especially if you keep them enclosed in the incubator too. I used to pierce my tape all the time for more GE and I never had contam. problems because of this reason. On the contrary, it always helped to get a faster move on.

This is terrible advice, If you poke holes in GE material, contamination will most certainly get in..


Your jars are contaminated with green mold trichoderma, start again and make sure you follow sterile procedure.

 

[Nereus]

I encourage you to take a sterile syringe needle and puncture the micropore tape a couple of times. Very small holes will still not let any airbornes or anything pass through, especially if you keep them enclosed in the incubator too. I used to pierce my tape all the time for more GE and I never had contam. problems because of this reason. On the contrary, it always helped to get a faster move on.

This is terrible advice, If you poke holes in GE material, contamination will most certainly get in..


Your jars are contaminated with green mold trichoderma, start again and make sure you follow sterile procedure.

Obviously you haven't ever tried this method. Why chime in, then, I wonder? Though I have no intention to argue, I dare to disagree, and tell you sir, that this practice has never yielded me nothing other than white healthy mycelium, strong healthy fruits, but not trichoderma, or even the slightest trace of metabolites what so ever. You can call it luck, I call it a very clean ,dust free environment, and a good sterile procedure overall... Also nobody said to "poke holes in GE material" There's a difference between poking ye needle holes and fine puncturing the micropore tape with a thin needle... yet again, tried and tested method, airborne proof :thumb_up:

 

[blue.magic]

I emptied the jars and 4 out of 6 smelled like rotten cheese, the mycelium turned greyish and I found the mould depicted the image.

But the remaining 2 jars were fine. I spawned them and they grow vigorously.

I used larger holes in the new jars and little bit more water and they colonize well.

The problem was most probably lack of AE.

 

[dmusicaltrancistor]

Hey Blue magic how do you find your incubator is working? I find when I try to use my incubator that i get too much condensation on the inside of my jars causing contams and jars to stall

have you noticed condensation occurring in your jars? If you did have this issue previously how did you prevent it from happening?
at the moment i have abandoned using my incubator until i get a better Tek for it, I just leave my jars to colonize at room temp.
I would like to get it working though to shorten colonization times

anyways good luck and grow them bad boys good